【摘要】 为改良柑桔品种,以普通溆浦甜橙(CitrussinensiscvXupu)实生苗上胚轴为外植体,建立了农杆菌介导的遗传转化体系,并把chit42基因和phyB基因导入其中.上胚轴经农杆菌工程菌液浸染后,在共培培养基(MS+3mg/LBA)上培养3d,然后分别转移到含有100mg/LKan(卡那霉素,Kanamycin)和500mg/LCef(头孢噻肟霉素,Cefotaxime)的选择培养基上培养14d后,上胚轴开始分化抗性芽,chit42基因和phyB基因的抗性芽再生率分别为27.8%和35.6%.将抗性芽在筛选培养基上培养30d后用茎尖微芽嫁接成苗.目前,嫁接苗在温室中生长良好,经PCR鉴定为转化植株.更多还原

【Abstract】 Seedling epicotyl(internodal stem) segments of ‘Xupu’ sweet orange(Citrus sinensis L.Osb.)were used as explants to establish Agrobacterium-mediated transformation system with chit42 and phyB genes.The results showed that transformed buds could be obtained after the epicotyl segments were cultured on co-culture medium(MS+3 mg/L BA) for 3 days and then on selective medium with 100 mg/L Kanamycin and 500 mg/L Cefotaxime for 2 weeks.The transformation efficency of Sweet orange with the chit42 and phyB genes reached 27.8% and 35.6%,respectively.After the regenerated buds were cultured for 4 weeks more,they were micro-grafted onto seedlings of Carrizo citrange grown in vitro and one month later regrafted on greenhouse grown rootstocks.Now the transgenic plants are growing well in the greenhouse,and the presence of chit42 and phyB genes in the regenerated plants were confirmed by PCR analysis.更多还原